摘要
利用硫酸铵盐析和柱层析对拟穴青蟹(Scylla paramamosain)进行处理,纯化得到28 ku的新型过敏原蛋白,经质谱鉴定为磷酸丙糖异构酶(triosephosphate isomerase,TIM),与中华绒螯(Eriocheir sinensis)TIM序列相似度达到93%。该过敏原蛋白能够与甲壳类过敏患者血清和兔抗克氏原螯虾多克隆抗体发生特异性反应,等电点为5.8。对热处理稳定,加热温度高于50℃时发生聚合,且多聚体仍具有免疫结合活性,p H>9.0时,对TIM免疫结合活性略有影响。在模拟胃肠液消化过程中,TIM耐受胰蛋白酶消化但不耐受胃蛋白酶消化,模拟胃液消化后的小分子片段仍保留有Ig G结合活性。综合血清学及性质分析,该磷酸丙糖异构酶是拟穴青蟹的一种新型过敏原。
A 28 ku protein was purified from crab（ Scylla paramamosain） and identified to be triosephosphate isomerase（ TIM） by mass spectrometry,which had a 93% similarity with the TIM sequence of Eriocheir sinensis and its isoelectric point was 5. 8. It also showed IgE-binding activity with shellfish allergic patients＇ sera and Ig G-binding activity with rabbit anti-Procambarus clarkii polyclonal antibody. TIM aggregated after being heated above 50 ℃ and the polymer remained allergenic reactivity. The binding activity of TIM reduced under strong alkaline condition（ p H 〉9. 0）. The allergen protein could be digested into small fragments by pepsin while it remained the IgG reactivity.
出处
《集美大学学报（自然科学版）》
CAS
2016年第6期428-437,共10页
Journal of Jimei University：Natural Science
基金
国家自然科学基金重点资助项目(U1405214
31171660)
关键词
拟穴青蟹
磷酸丙糖异构酶
新型过敏原
纯化
性质分析
Scylla paramamosain
triosephosphate isomerase
novel allergen
purification
physico chemical characterization
